In vitro and in vivo hepatic transport of the magnetic resonance imaging contrast agent B22956/1: role of MRP proteins

doi:10.1016/S0006-291X(02)00182-1

Biochemical and Biophysical Research Communications

Volume 293, Issue 1, 26 April 2002, Pages 100-105

https://doi.org/10.1016/S0006-291X(02)00182-1 Get rights and content

Abstract

The molecular mechanisms of the hepatic transport of B22956/1, a new gadolinium complex from the class of intravascular contrast agents for MRI, which undergoes extensive biliary elimination, were studied. Biliary and urinary elimination of B22956/1 were measured in normal and in mutant MRP2 lacking rats (TR⁻); cellular trafficking of the compound was assessed in wild and MRP1 or MRP2 transfected MDCKII cells. Eight hours after IV injection of B22956/1, 90±8% of the dose was recovered in the bile of normal rats. By contrast, in TR⁻ rats, the biliary excretion was significantly lower (14±3%) while 55±9% of the compound was found in urine. In vitro, the cellular accumulation of B22956/1 was significantly lower in both MRP1 and MRP2 transfected cells as compared to wild type MDCKII cells, and the cellular efflux was prevented by the MRP inhibitor MK571, indicating the involvement of both MRP2 and MRP1 in the transport of B22956/1. Due to the distinct cellular localization of the proteins, MRP2 accounts for the biliary and urinary excretion of the compound, while MRP1 prevents cellular accumulation of the MRI agent. B22956/1 may be useful in clinical conditions where a defective biliary transport is present.

Section snippets

Materials and methods

Materials. B22956/1 is the trisodium salt of a derivative of gadopentetate bearing on the methylene group of the centrally located acetate group in S-configuration, a propionic acid linker to the amino group of the 3β-amino-analogue of deoxycholic acid [9]. Its chemical name according to CAS is trisodium[(3β,5β,12α)-3-[[(4S)-4-[bis[2-[bis[(carboxy-κO)methyl]amino-κN]ethyl]amino-κN]-4-(carboxy-κO)-1-oxybutyl]amino]-12-hydroxycholan-24-oato (6-)]gadolinate (3-) and the proposed international

“In vivo” studies

The overall biliary excretion of gadolinium, expressed as percentage of the administered dose, is reported in Table 1. In control rats, 480 min after intravenous administration of B22956/1, the biliary excretion of gadolinium accounted for 89.6±8.8% of the injected dose. On the contrary, the biliary excretion was significantly lower in the TR⁻ rats, where the total gadolinium recovery in bile was 14.1±3.0% (p<0.001). In control rats the biliary excretion was almost maximal after 120 min when

Discussion

Many attempts have been performed to identify contrast agents which may show organ specificity and thus may be “targeted” to specific organs. As far as the study of hepatobiliary MRI is concerned, data have been provided that several Gd-complex MRI contrast agents way undergo an efficient hepatic uptake and subsequent biliary excretion. While the molecular mechanisms involved in the first step are still not fully unraveled, evidence has been provided indicating a role of MRP proteins in the

Acknowledgements

We thank Dr. R.Oude Elferink for the kind supply of MRP1 and MRP2 stably transfected MDCKII cells. LP and CF were supported by a career development grant by Bracco SpA. The work was supported in part by an intramural research grant (60%-00) and by a grant from Fondo Studio Malattie Fegato (CRT-CSF/00/1).

References (24)

L. Pascolo et al.
Abc protein transport of MRI contrast agents in canalicular rat liver plasma vesicles and yeast vacuoles
Biochem. Biophys. Res. Commun.
(2001)
C. de Haen et al.
Hepatic transport of gadobenate dimeglumine in TR-rats
Acad. Radiol.
(1996)
L. Pascolo et al.
Mechanisms for the transport of unconjugated bilirubin in human trophoblastic BeWo cells
FEBS Lett.
(2001)
I. Leier et al.
ATP-dependent para-aminohippurate transport by apical multidrug resistance protein MRP2
Kidney Int.
(2000)
M. Buchler et al.
cDNA cloning of the hepatocyte canalicular isoform of the multidrug resistance protein, cMrp, reveals a novel conjugate export pump deficient in hyperbilirubinemic mutant rats
J. Biol. Chem.
(1996)
J. Kartenbeck et al.
Absence of the canalicular isoform of the MRP gene-encoded conjugate export pump from the hepatocytes in Dubin–Johnson syndrome
Hepatology
(1996)
P. Borst et al.
The multidrug resistance protein family
Biochim. Biophys. Acta
(1999)
E. Bakos et al.
Functional multidrug resistance protein (MRP1) lacking the N-terminal transmembrane domain
J. Biol. Chem.
(1998)
F.M. Cavagna, J. Zheng, V. Lorusso, F. Maggioni, D. Li, P.J. Finn, M.R. coronary angiographic: new insight and new...
F.M. Cavagna, J. Zheng, V. Lorusso, F. Maggioni, D. Li, P.J. Finn, New protein binding gadolinium chelate with high...

M. Muller et al.

Complete elimination of the hepatobiliary MR contrast agents in hepatic disfunction: an experimental study using transport-deficient, mutant rats

MAGMA

(1994)

P.L. Jansen et al.

Selective hepatobiliary transport defect for organic anions and neutral steroids in mutant rats with hereditary-conjugated hyperbilirubinemia

Hepatology

(1987)

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In vitro and in vivo hepatic transport of the magnetic resonance imaging contrast agent B22956/1: role of MRP proteins

Abstract

Section snippets

Materials and methods

“In vivo” studies

Discussion

Acknowledgements

Biochem. Biophys. Res. Commun.

Acad. Radiol.

FEBS Lett.

Kidney Int.

J. Biol. Chem.

Hepatology

Biochim. Biophys. Acta

J. Biol. Chem.

Complete elimination of the hepatobiliary MR contrast agents in hepatic disfunction: an experimental study using transport-deficient, mutant rats

MAGMA

Selective hepatobiliary transport defect for organic anions and neutral steroids in mutant rats with hereditary-conjugated hyperbilirubinemia

Hepatology